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포플러의 미성숙 배와 캘러스에서 유전자총에 의한 GUS - gene 의 일시적 발현
강호덕,강상구,배한홍,박교수,( Ho Duck Kang,Sang Gu Kang,Han Hong Bae,Kyo Soo Park,Richard B . Hall ) 한국산림과학회 1997 한국산림과학회지 Vol.86 No.3
Excised immature ovules and calli derived from the stems of cottonwood were bombarded with microprojectiles carrying plasmid DNA containing CaMV-35S promoter and β-glucuronidase(GUS) gene. After bombarded, the expression of GUS gene was detected by the assay of 5-bromo-4-chloro-3-indolyl-β-gluconide(X-gluc). Transient gene expression was measured by counting the number of distinct regions of GUS activity per explant. As major parameters, the number of shots and the period of exposure to X-gluc after the bombardment were investigated for detecting GUS gene expression. In this experiment, the percents of GUS gene expression showing spots were 56.8 from immature ovules and 75.9 from micro-calli of cottonwood species. Among the treatments, two consecutive shots and 48 hour exposure produced abet 25.75±2.77(per ovule), 11.43±1.22(per mini petridish) spots, respectively, Microprojectile particle bombardment provides a useful method to assay transient expression in both types of explants. Furthermore, our results represent that the excised ovule and/or the calli might be stably transformed by the biolistics.